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Microbial Metabolism

Chapter 4
Objectives
Define metabolism and the processes involved in
it
Cite the role of enzymes and energy in
metabolism
Classify enzymes according to activity
Discuss ATP
Discuss the different metabolic pathways in
glucose catabolism
Identify and explain the three processes involved
in aerobic respiration
Metabolism sum of all chemical
processes that occur within a cell.
1. Catabolism ( Catabolic )
breakdown of complex organic molecules into simpler compounds
releases ENERGY
2. Anabolism ( Anabolic )
the building of complex organic molecules from simpler ones
requires ENERGY
Catabolism and Anabolism
Catabolism Anabolism

All of the catabolic reactions in cell All of the anabolic reactions in the cell

Catabolic reactions release energy Anabolic reactions require energy

Involve the breaking of bonds; Involve in the creation of bonds; it


whenever chemical bonds are broken, takes energy to create chemical bonds
energy is released

Larger molecules are broken down into Smaller molecules are bonded
smaller molecules (sometimes referred together to create larger molecules
to as a degradative reactions) (sometimes referred to as biosynthetic
reactions)
Enzymes - are special group of protein
molecules that bring about a chemical change
while themselves remaining unchanged.
- Catalysts that speed chemical reactions.

A. Enzymes are substrate specific


Lipases Lipids
Sucrases Sucrose
Ureases Urea
Proteases Proteins
DNases DNA
Substrate - Substance acted upon by enzyme
End products products of enzyme reactions
Characteristics of an Enzymes
Catalyze or speed up chemical processes
Reusable
Enzyme activity is highly specific
Sensitive to any physical or chemical agents
Enzyme Specificity can be explained
by the Lock and Key Theory

E + S -----> ES ------> E + P
Types of enzymes according to their activity

1. Hydrolases operate in hydrolysis reactions where


the products of water are added to the end products.

example: Lipase and peptidase

2. Oxidases catalyzed oxidation-reduction reactions.


3. Transferases operate in transfer reactions.

example: kinases, transaminases


Naming of Enzymes - most are named
by adding ase to the substrate

Sucrose Sucrase
Lipids Lipase
DNA DNase
Proteins Protease
removes a Hydrogen Dehydrogenase
removes a phosphate phosphotase
Naming of Enzymes
Grouped based on type of reaction they catalyze
1. Oxidoreductases oxidation & reduction
2. Hydrolases hydrolysis
3. Ligases synthesis
Enzyme Components
2 Parts
1. Apoenzyme - protein portion
2. Coenzyme (cofactor) - non-protein

Holoenzyme - whole enzyme


Coenzymes
Many are derived from vitamins

1. Niacin
NAD (Nicotinamide adenine dinucleotide)
2. Riboflavin
FAD (Flavin adenine dinucleotide)
3. Pantothenic Acid
CoEnzyme A
Factors that Influence Enzymatic
Activity

Denaturation of an Active Protein


The rate of an enzyme controlled reaction is
affected by temperature
At low temperatures enzyme controlled reactions go slowly
because the molecules have low kinetic energy.
When temperature increases the reaction also increases as the
molecules have more kinetic energy

But this only occurs up to the


optimum temperature (usually
about 40oC)

The temperature at which


the rate of reaction is
fastest is known as the
optimum temperature
After the optimum temperature the heat causes the
enzyme to denature

The enzyme changes shape and the active site no longer matches
the shape of the substrate molecule
Rate of reaction of an enzyme reaction changes at
different temperatures

Optimum temperature

Molecules gain Enzyme


Rate kinetic energy is denaturing
Of
Reaction

0 10 20 30 40 50 60 70
Temperature/oC
The activity and shape of enzymes is also
affected by pH
Enzymes prefer to work at an optimum pH. Outside of its pH range
the enzyme is denatured.

Optimum pH
pepsin amylase

Rate
Of
Reaction

1 2 3 4 5 6 7 8 9 10 11 12
pH
Inhibitors can effect enzymatic activity
1. Competitive Inhibitors
2. Noncompetitive Inhibitors
Competitive Inhibitors -compete for the
active site
1. Penicillin
competes for the active site on the enzyme involved in the synthesis of
the pentaglycine crossbridge
2. Sulfanilamide (Sulfa Drugs)
competes for the active site on the enzyme that converts PABA into
Folic Acid
Folic Acid - required for the synthesis of DNA and RNA
Non-competitive Inhibitors - attach to
an allosteric site
Energy and ATP

A highly energy yielding compound which


supplements enzymes in the biochemical
reactions of metabolism
Moves to any part of the cell where energy
consuming reactions is taking place and serve
as energy source
In bacterium, ATP supplies energy for binary fission,
flagellar motion and spore formation
It fuels protein synthesis and carbohydrate breakdown
Carbohydrate Catabolism
Microorganisms oxidize carbohydrates as their primary source
of energy
Glucose - most common energy source
Energy obtained from Glucose by:
Respiration
Fermentation
Aerobic Cellular Respiration

Electrons released by oxidation are passed down an Electron


Transport System with oxygen being the Final Electron Acceptor

General Equation:

Glucose + oxygen----> Carbon dioxide + water


ATP
Chemical Equation
C6H12O6 + 6 O2 -------> 6 CO2 + 6 H2O

38 ADP + 38 P 38 ATP
Aerobic Cellular Respiration
4 subpathways

1. Glycolysis
2. Transition Reaction
3. Krebs Cycle
4. Electron Transport System
1. Glycolysis (splitting of sugar)
Oxidation of Glucose into 2 molecules of Pyruvic acid
Embden-Meyerhof Pathway-major route of glucose
metabolism
Occurs in the cytoplasm of bacteria
End Products of Glycolysis:
2 Pyruvic acid
2 NADH2
2 ATP
2. Transition Reaction
Connects Glycolysis to Krebs Cycle

End Products:
2 Acetyl CoEnzyme A
2 CO2
2 NADH2
3. Krebs Cycle (Citric Acid Cycle)
Series of chemical reactions that begin and end with citric acid
Occurs in the cell membrane

Products:
2 ATP
6 NADH2
2 FADH2
4 CO2
4. Electron Transport System
Occurs within the cell membrane of Bacteria

Chemiosomotic Model of Mitchell


34 ATP
How 34 ATP from E.T.S. ?
3 ATP for each NADH2
2 ATP for each FADH2

NADH2 FADH2

Glycolysis 2 Glycolysis 0
T. R. 2 T.R. 0
Krebs Cycle 6 Krebs Cycle 2

Total 10 Total 2

10 x 3 = 30 ATP 2 x 2 = 4 ATP
Total ATP production for the
complete oxidation of 1 molecule
of glucose in Aerobic Respiration
ATP
Glycolysis 2
Transition Reaction 0
Krebs Cycle 2
E.T.S. 34

Total 38 ATP
Anaerobic Respiration
Electrons released by oxidation are passed down an E.T.S., but
oxygen is not the final electron acceptor

Nitrate (NO3-) ----> Nitrite (NO2-)


Sulfate (SO24-) ----> Hydrogen Sulfide (H2S)
Carbonate (CO24-) -----> Methane (CH4)
Fermentation
Anaerobic process that does not use the E.T.S. Usually
involves the incomplete oxidation of a carbohydrate which
then becomes the final electron acceptor.

Glycolysis - plus an additional step


Fermentation may result in numerous
end products

1. Type of organism
2. Original substrate
3. Enzymes that are present and active
1. Lactic Acid Fermenation
Only 2 ATP
End Product - Lactic Acid
Food Spoilage
Food Production
Yogurt - Milk
Pickles - Cucumbers
Sauerkraut - Cabbage
2 Genera:
Streptococcus
Lactobacillus
2. Alcohol Fermentation
Only 2 ATP
End products:
alcohol
CO2
Alcoholic Beverages

Saccharomyces cerevisiae (Yeast)


Propionic Acid Fermentation
Only 2 ATP
End Products:
Propionic acid
CO2

Propionibacterium sp.
Fermentation End Products

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